title: In vitro generation of mature, naive antigen-specific CD8(+) T cells with a single T-cell receptor by agonist selection.
abstract: Peripheral blood T cells transduced with a tumor-specific T-cell receptor (TCR) face problems of auto-reactivity and lack of efficacy caused by cross-pairing of exogenous and endogenous TCR chains, as well as short term in vivo survival due to activation and growth factor-induced differentiation. We here studied an alternative strategy for the efficient generation of naive CD8(+) T cells with a single TCR. TCR-transduced human postnatal thymus-derived and adult mobilized blood-derived hematopoietic progenitor cells (HPCs) were differentiated to CD4(+)CD8(+) double-positive T cells using OP9-Delta-like 1 (OP9-DL1) cultures. Addition of the agonist peptide induced double positive cells to cross-present the peptide, leading, in the absence of co-stimulation, to cell cycle arrest and differentiation into mature CD8(+) T cells. Comprehensive phenotypic, molecular and functional analysis revealed the generation of naive and resting CD8(+) T cells through a process similar to thymic positive selection. These mature T cells show a near complete inhibition of endogenous TCRA and TCRB rearrangements and express high levels of the introduced multimer-reactive TCR. Upon activation, specific cytokine production and efficient killing of tumor cells were induced. Using this strategy, large numbers of high-avidity tumor-specific naive T cells can be generated from readily available HPCs without TCR chain cross-pairing.
authors.affiliation: Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, Ghent, Belgium.
authors.forename: S
authors.initials: S
authors.lastname: Snauwaert
authors.affiliation: Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, Ghent, Belgium.
authors.forename: G
authors.initials: G
authors.lastname: Verstichel
authors.affiliation: Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, Ghent, Belgium.
authors.forename: S
authors.initials: S
authors.lastname: Bonte
authors.affiliation: Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, Ghent, Belgium.
authors.forename: G
authors.initials: G
authors.lastname: Goetgeluk
authors.affiliation: Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, Ghent, Belgium.
authors.forename: S
authors.initials: S
authors.lastname: Vanhee
authors.affiliation: Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, Ghent, Belgium.
authors.forename: Y
authors.initials: Y
authors.lastname: Van Caeneghem
authors.affiliation: Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, Ghent, Belgium.
authors.forename: K
authors.initials: K
authors.lastname: De Mulder
authors.affiliation: Laboratory of Molecular & Cellular Therapy of the Department of Physiology and Immunology, Free University Brussels (VUB), Brussels, Belgium.
authors.forename: C
authors.initials: C
authors.lastname: Heirman
authors.affiliation: Department of Immunology, University College London, London, UK.
authors.forename: H
authors.initials: H
authors.lastname: Stauss
authors.affiliation: Laboratory of Experimental Hematology of the Department of Hematology, Leids Universitair Medisch Centrum, Leiden, The Netherlands.
authors.forename: M H M
authors.initials: MH
authors.lastname: Heemskerk
authors.affiliation: Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, Ghent, Belgium.
authors.forename: T
authors.initials: T
authors.lastname: Taghon
authors.affiliation: Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, Ghent, Belgium.
authors.forename: G
authors.initials: G
authors.lastname: Leclercq
authors.affiliation: Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, Ghent, Belgium.
authors.forename: J
authors.initials: J
authors.lastname: Plum
authors.affiliation: Department of Immunology, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: A W
authors.initials: AW
authors.lastname: Langerak
authors.affiliation: Laboratory of Molecular & Cellular Therapy of the Department of Physiology and Immunology, Free University Brussels (VUB), Brussels, Belgium.
authors.forename: K
authors.initials: K
authors.lastname: Thielemans
authors.affiliation: Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, Ghent, Belgium.
authors.forename: T
authors.initials: T
authors.lastname: Kerre
authors.affiliation: Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, Ghent, Belgium.
authors.forename: B
authors.initials: B
authors.lastname: Vandekerckhove
mesh_terms: D000328:Adult; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002648:Child; D002675:Child, Preschool; D015329:Gene Rearrangement, T-Lymphocyte; D006801:Humans; D016219:Immunotherapy, Adoptive; D007223:Infant; D007231:Infant, Newborn; D011948:Receptors, Antigen, T-Cell / Q000819:agonists / Q000502:physiology*
chemical_list: D011948:Receptors, Antigen, T-Cell